Dynamic culture yields engineered myocardium with near-adult functional output.
Dynamic culture yields engineered myocardium with near-adult functional output.
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DOI：
10.1016/j.biomaterials.2016.09.024
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发表时间：
2016-12
期刊：
影响因子：
14
通讯作者：
Bursac, Nenad
中科院分区：
文献类型：
--
作者：
Jackman, Christopher P.;Carlson, Aaron L.;Bursac, Nenad
关键词：
工程心脏组织有望用于细胞治疗和药物开发，但表现出不足的功能和成熟度。在这项研究中，我们试图显着提高大鼠和人类工程心脏组织的功能和成熟。我们开发了用于工程化"心脏束"的动态自由浮动培养条件，所述心脏束是由包埋在纤维蛋白基水凝胶中的新生大鼠心肌细胞或人多能干细胞衍生的心肌细胞（hPSC-CM）制成的三维圆柱形组织。与静态培养相比，2周动态培养的新生大鼠心肌束显著增加了肌节蛋白的表达、心肌细胞大小（~2.1倍）、收缩力（~3.5倍）和动作电位传导速度（~1.4倍）。平均收缩力/横截面积（59.7 mN/mm2）和传导速度（52.5 cm/sec）匹配或接近成年大鼠心肌，分别。静态培养的心束的劣功能被转移到动态条件下，这是伴随着mTORC 1活性的增加和AMPK磷酸化的下降，并被雷帕霉素阻断救援。此外，动态培养效应不刺激ERK 1/2通路，对机械敏感通道的阻断剂不敏感，这表明增加营养的可用性，而不是机械刺激作为mTORC1的上游激活剂。与苯乙哌啶治疗的直接比较证实，动态培养促进了生理性心肌细胞生长，而不是病理性肥大。优化的动态培养条件还增强了由源自多个hPSC系的心肌细胞可重复地制备的人心束的功能，导致收缩力（~2.5倍）和传导速度（~1.4倍）显著增加。平均比力为23.2 mN/mm2，传导速度为25.8 cm/sec，接近成人心肌的功能指标。总之，我们已经开发了一种通用的方法，用于工程化心脏组织，具有接近成人的功能输出，而不需要外源性电刺激或机械刺激，并已确定mTOR信号传导作为体外促进组织成熟和功能的重要机制。
Engineered cardiac tissues hold promise for cell therapy and drug development, but exhibit inadequate function and maturity. In this study, we sought to significantly improve the function and maturation of rat and human engineered cardiac tissues. We developed dynamic, free-floating culture conditions for engineering “cardiobundles”, 3-dimensional cylindrical tissues made from neonatal rat cardiomyocytes or human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) embedded in fibrin-based hydrogel. Compared to static culture, 2-week dynamic culture of neonatal rat cardiobundles significantly increased expression of sarcomeric proteins, cardiomyocyte size (~2.1-fold), contractile force (~3.5-fold), and conduction velocity of action potentials (~1.4-fold). The average contractile force per cross-sectional area (59.7 mN/mm2) and conduction velocity (52.5 cm/sec) matched or approached those of adult rat myocardium, respectively. The inferior function of statically cultured cardiobundles was rescued by transfer to dynamic conditions, which was accompanied by an increase in mTORC1 activity and decline in AMPK phosphorylation and was blocked by rapamycin. Furthermore, dynamic culture effects did not stimulate ERK1/2 pathway and were insensitive to blockers of mechanosensitive channels, suggesting increased nutrient availability rather than mechanical stimulation as the upstream activator of mTORC1. Direct comparison with phenylephrine treatment confirmed that dynamic culture promoted physiological cardiomyocyte growth rather than pathological hypertrophy. Optimized dynamic culture conditions also augmented function of human cardiobundles made reproducibly from cardiomyocytes derived from multiple hPSC lines, resulting in significantly increased contraction force (~2.5-fold) and conduction velocity (~1.4-fold). The average specific force of 23.2 mN/mm2 and conduction velocity of 25.8 cm/sec approached the functional metrics of adult human myocardium. In conclusion, we have developed a versatile methodology for engineering cardiac tissues with a near-adult functional output without the need for exogenous electrical or mechanical stimulation, and have identified mTOR signaling as an important mechanism for advancing tissue maturation and function in vitro.
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影响因子：
10.9
作者：
Howell, Jessica J.;Manning, Brendan D.
通讯作者：
Manning, Brendan D.
影响因子：
9.5
作者：
Hirt MN;Sörensen NA;Bartholdt LM;Boeddinghaus J;Schaaf S;Eder A;Vollert I;Stöhr A;Schulze T;Witten A;Stoll M;Hansen A;Eschenhagen T
通讯作者：
Eschenhagen T
影响因子：
48
作者：
Burridge, Paul W.;Matsa, Elena;Shukla, Praveen;Lin, Ziliang C.;Churko, Jared M.;Ebert, Antje D.;Lan, Feng;Diecke, Sebastian;Huber, Bruno;Mordwinkin, Nicholas M.;Plews, Jordan R.;Abilez, Oscar J.;Cui, Bianxiao;Gold, Joseph D.;Wu, Joseph C.
通讯作者：
Wu, Joseph C.
影响因子：
--
作者：
BOLTER, CP;ATKINSON, KJ
通讯作者：
ATKINSON, KJ
影响因子：
4.1
作者：
Boudou, Thomas;Legant, Wesley R.;Chen, Christopher S.
通讯作者：
Chen, Christopher S.